ABSTRACT
The objective of this work was to identify a human use-permissible adjuvant to enhance significantly the antibody response to a recombinant anti-hCG vaccine. Previous Phase II efficacy trials in sexually active women have demonstrated the prevention of pregnancy at hCG bioneutralization titers of 50ng/ml or more. Mycobacterium indicus pranii (MIP), a non-pathogenic Mycobacterium employed as an autoclaved suspension in aqueous buffer, significantly increased antibody titers in the FVB strain of mice. Three other genetic strains of mice: SJL, C3H, and C57Bl/6 responded with antibody titers several-fold higher than 50 ng/ml, which is the protective threshold in women, although there were differences in the peak titers attained. In addition, the duration of the antibody response was lengthened. The vaccine hCGß-LTB, given together with MIP, induces both a Th1 and Th2 response, which is reflected in the production of not only IgG1, but also a high proportion of IgG2a and IgG2b antibodies.
Subject(s)
Antibodies/immunology , Chorionic Gonadotropin/immunology , Contraceptive Agents, Female/immunology , Immunologic Factors/immunology , Mycobacterium/immunology , Animals , Antibodies/blood , Chorionic Gonadotropin/pharmacology , Clinical Trials, Phase II as Topic , Contraceptive Agents, Female/pharmacology , Female , Humans , Immunologic Factors/pharmacology , Mice , Mice, Inbred C3H , Vaccines, Synthetic/immunology , Vaccines, Synthetic/pharmacologyABSTRACT
Human chorionic gonadotropin (hCG) is synthesized soon after fertilization and is essential for embryonic implantation. A vaccine targeting hCG would be an ideal choice for immuno-contraception; an anti-hCG vaccine developed by Talwar et al., has previously undergone Phase II efficacy trials, providing proof of principle. These trials established the threshold levels of bio-neutralizing anti-hCG antibody titers required to prevent pregnancy; however, these titers (>50 ng/ml) were achieved in only 80% of immunized women. In this communication, we report a novel recombinant anti-hCG vaccine which demonstrates improved immunogenicity. hCGß was genetically fused at C-terminal to the B-subunit of E. coli heat-labile enterotoxin. The recombinant fusion protein (hCGß-LTB) was expressed in Pichia pastoris and, upon adsorption on Alhydrogel along with Mycobacterium indicus pranii (MIP) as an immuno-modulator, evoked a very high anti-hCG immune response in 100% of immunized BALB/c mice. This recombinant vaccine is expected to reduce cost as well as facilitate production of a molecularly consistent conjugate on a large scale.
Subject(s)
Chorionic Gonadotropin, beta Subunit, Human/immunology , Vaccines, Contraceptive/immunology , Animals , Antibodies, Neutralizing/blood , Chorionic Gonadotropin, beta Subunit, Human/genetics , Chorionic Gonadotropin, beta Subunit, Human/isolation & purification , Cloning, Molecular , Female , Humans , Mice , Mice, Inbred BALB C , Pichia/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Vaccines, Synthetic/immunologyABSTRACT
Gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) are unique targets for the control of fertility. Immunological approaches to neutralizing these hormones have additional utility in cancer treatment. Vaccines have been developed against both GnRH and hCG and these have undergone Phase I/II clinical trials documenting their safety, reversibility and efficacy. The heterospecies dimer hCG vaccine prevented pregnancy in women of proven fertility without impairment of ovulation or derangement of menstrual regularity and bleeding profiles. The protective threshold of antibody titers to achieve efficacy was determined in these first-ever trials. Recently, a recombinant vaccine against the beta subunit of hCG linked to the B subunit of heat labile enterotoxin has been made and expressed as a glycosylated conjugate in Pichia pastoris. Experiments indicate its ability to generate antibodies above the protective threshold in all immunized Balb/c mice. Ectopic expression of hCG/hCGbeta is observed in many advanced stage cancers of various origins. A chimeric high affinity and specific recombinant antibody against hCGbeta linked to curcumin kills hCGbeta expressing T lymphoblastic leukemia cells without any deleterious effect. Several synthetic and recombinant vaccines have been developed against GnRH. These reduce serum testosterone to castration levels causing atrophy of the prostate. Three Phase I/II clinical trials conducted in India and Austria have shown that these vaccines elicit non-surgical reduction of testosterone, a fall in prostate specific antigen and clinical improvement of prostate carcinoma patients. A multimer recombinant vaccine against GnRH has high efficacy for sterilization of pigs and other animals.
